Rapid detection of clonal T-cell receptor-β gene rearrangements in T-cell lymphomas using the lightCycler-polymerase chain reaction with DNA melting curve analysis

Xiao Yang Yang, Dongsheng Xu, Juan Du, Hideko Kamino, Jennifer Rakeman, Howard Ratech

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Various molecular methods have been developed to diagnose clonal T-cell receptor (TCR) gene rearrangements in clinical samples. Most polymerase chain reaction strategies for detecting clonal TCR gene rearrangements rely on either gel or capillary electrophoresis. However, a cumbersome manual transfer step separates amplification from analysis. Recently, we developed a novel polymerase chain reaction assay using the LightCycler system to detect clonal immunoglobulin heavy chain gene rearrangement. In the current study, we extend this work to include the TCR. We report that clonal TCR-β (TCR-β) gene rearrangements can be detected in less than 1 hour after preparing the DNA by measuring DNA melting immediately after amplification in a single closed capillary tube. We retrospectively studied 52 fresh-frozen tissue samples from patients clinically suspected of T-cell malignancy. A clonal TCR-β gene rearrangement was detected in 14 samples by DNA melting curve analysis. When DNA melting was compared to the gold standard methods of Southern blot or denaturing gradient gel electrophoresis, it achieved a sensitivity equal to 71% and a specificity equal to 94%. We also compared melting curve analysis and polyacrylamide gel electrophoresis: melting curve analysis reached a sensitivity equal to 100% and a specificity equal to 97%. We conclude that DNA melting curve analysis in the LightCycler system has potential for clinical use as a new, ultra-fast method for the initial diagnosis of clonal TCR-β gene rearrangements.

Original languageEnglish (US)
Pages (from-to)81-88
Number of pages8
JournalJournal of Molecular Diagnostics
Volume7
Issue number1
StatePublished - Feb 2005

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T-Lymphocyte Gene Rearrangement
Nucleic Acid Denaturation
T-Cell Receptor Genes
T-Cell Lymphoma
Polymerase Chain Reaction
T-Cell Antigen Receptor
Freezing
Immunoglobulin Heavy Chain Genes
Denaturing Gradient Gel Electrophoresis
Gene Rearrangement
Capillary Electrophoresis
Southern Blotting
Polyacrylamide Gel Electrophoresis
Gels
T-Lymphocytes
DNA
Neoplasms

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Rapid detection of clonal T-cell receptor-β gene rearrangements in T-cell lymphomas using the lightCycler-polymerase chain reaction with DNA melting curve analysis. / Yang, Xiao Yang; Xu, Dongsheng; Du, Juan; Kamino, Hideko; Rakeman, Jennifer; Ratech, Howard.

In: Journal of Molecular Diagnostics, Vol. 7, No. 1, 02.2005, p. 81-88.

Research output: Contribution to journalArticle

Yang, Xiao Yang ; Xu, Dongsheng ; Du, Juan ; Kamino, Hideko ; Rakeman, Jennifer ; Ratech, Howard. / Rapid detection of clonal T-cell receptor-β gene rearrangements in T-cell lymphomas using the lightCycler-polymerase chain reaction with DNA melting curve analysis. In: Journal of Molecular Diagnostics. 2005 ; Vol. 7, No. 1. pp. 81-88.
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